Psma targeted fluorescent agents for image guided surgery

ABSTRACT

Compositions and methods for visualizing tissue under illumination with near-infrared radiation, including compounds comprising near-infrared, closed chain, sulfo-cyanine dyes and prostate specific membrane antigen ligands are disclosed.

CROSS-REFERENCE TO RELATED APPLICATIONS

This application claims the benefit of U.S. Provisional Application No.62/324,097, filed Apr. 18, 2016, and is a continuation-in-part of U.S.patent application Ser. No. 14/243,535, filed Apr. 2, 2014, which is adivisional of U.S. patent application Ser. No. 13/257,499 filed Sep. 19,2011, and now U.S. Pat. No. 9,056,841 issued Jun. 16, 2015, which is a35 U.S.C. §371 National Stage Entry of International Application No.PCT/US2010/028020 having an international filing date of Mar. 19, 2010,which claims the benefit of U.S. Provisional Application No. 61/248,934filed Oct. 6, 2009, U.S. Provisional Application No. 61/248,067 filedOct. 2, 2009, U.S. Provisional Application No. 61/161,484 filed Mar. 19,2009, and U.S. Provisional Application No. 61/161,485 filed Mar. 19,2009, each of which is incorporated herein by reference in its entirety.

STATEMENT OF GOVERNMENTAL INTEREST

This invention was made with U.S. government support under grant no.CA134675 to the National Institutes of Health (NIH). The U.S. governmenthas certain rights in the invention.

BACKGROUND

Prostate cancer (PCa) is the most commonly diagnosed malignancy and thesecond leading cause of cancer-related death in men in the UnitedStates. Only one half of tumors due to PCa are clinically localized atdiagnosis and one half of those represent extracapsular spread.Localization of that spread, as well as determination of the total bodyburden of PCa, has important implications for therapy.

Prostate-specific membrane antigen (PSMA) is a marker forandrogen-independent disease that also is expressed on solid(nonprostate) tumor neovasculature. Complete detection and eradicationof primary tumor and metastatic foci are required to effect a cure inpatients with cancer; however, current preoperative assessment oftenmisses small metastatic deposits. Accordingly, more sensitive imagingtechniques are required, including those that can allow visualization ofthe tumor during surgery.

SUMMARY

In some aspects, the presently disclosed subject matter provides thefollowing compound:

or a pharmaceutically acceptable salt thereof.

In other aspects, the presently disclosed subject matter provides acomposition comprising compound (3), as provided immediatelyhereinabove, wherein the composition is adapted for visualization oftissue under illumination with near-infrared radiation. In certainaspects, the composition is adapted for administration to a subject. Inyet more certain aspects, the composition comprises a unit dosage formof compound (3). In particular aspects, the unit dosage form delivers tothe subject an amount of compound (3) between about 0.01 and about 8mg/kg. In more particular aspects, the unit dosage form delivers to thesubject an amount of compound (3) of about 0.01 mg/kg, about 0.05 mg/kg,about 0.10 mg/kg, about 0.20 mg/kg, about 0.3 mg/kg, about 0.35 mg/kg,about 0.40 mg/kg, about 0.45 mg/kg, about 0.50 mg/kg, about 0.55 mg/kg,about 0.60 mg/kg, about 0.65 mg/kg, about 0.70 mg/kg, about 0.75 mg/kg,about 0.80 mg/kg, about 0.90 mg/kg, about 1 mg/kg, about 2, mg/kg, about4 mg/kg, about 6 mg/kg, or about 8 mg/kg. In particular aspects, thecomposition is in a single dose form.

In certain aspects, the composition is in dry form. In more certainaspects, the composition is lyophilized in a sterile container. Inparticular aspects, the composition is contained within a sterilecontainer. In yet more particular aspects, the sterile containercomprises a machine detectable identifier.

In some aspects, the composition further comprises one or morepharmaceutically acceptable excipients in an oral dosage form. In otheraspects, the composition further comprises one or more pharmaceuticallyacceptable carriers in an injectable dosage form. In certain aspects,the composition further comprises one or more pharmaceuticallyacceptable excipients in a dosage form for direct delivery to a surgicalsite.

In other aspects, the presently disclosed subject matter provides forthe use of a composition comprising compound (3) for administration to asubject to obtain visualization of tissue expressing PSMA underillumination with near-infrared radiation. In certain aspects, thesubject is a human subject.

In other aspects, the presently disclosed subject matter provides amethod for visualization of tissue expressing PSMA, the methodcomprising administering to a subject a composition comprising compound(3), wherein compound (3) is administered in an amount sufficient forimaging tissue under illumination with near-infrared radiation; imagingthe tissue under illumination with near-infrared radiation; andobtaining at least one image of tissue from the subject.

In certain aspects, the composition comprises a unit dosage form ofcompound (3). In more certain aspects, the unit dosage form delivers tothe subject an amount of compound (3) from about 0.01 mg/kg and about 8mg/kg. In particular aspects, the composition is sterile, non-toxic, andadapted for administration to a subject.

In certain aspects, the method further comprises obtaining the imageduring administration, after administration, or both during and afteradministration of the composition. In other aspects, the method furthercomprises intravenously injecting a composition comprising compound (3)into the subject. In particular aspects, the composition is injectedinto a circulatory system of the subject.

In certain aspects, the method further comprises visualizing a subjectarea on which surgery is or will be performed. In more certain aspects,the method further comprises performing a surgical procedure of thesubject area based on the visualization of the area. In yet more certainaspects, the method further comprises viewing a subject area on which anophthalmic, arthroscopic, laparoscopic, cardiothoracic, muscular, orneurological procedure is or will be performed.

In certain aspects, the method further comprises diagnosing the subjectwith a condition or disease based on the visualization of the tissueexpressing PSMA. In more certain aspects, the method further comprisesobtaining ex vivo images of at least a portion of the subject. Inparticular aspects, the tissue being visualized comprises tumor tissue.In more particular aspects, the tissue being visualized comprisescancerous tissue. In even more particular aspects, the tissue beingvisualized comprises prostate tissue. In even yet more particularaspects, the tissue being visualized comprises prostate tumor tissue. Inother aspects, the tissue being visualized comprises nerve tissue.

Certain aspects of the presently disclosed subject matter having beenstated hereinabove, which are addressed in whole or in part by thepresently disclosed subject matter, other aspects will become evident asthe description proceeds when taken in connection with the accompanyingExamples and FIGURE as best described herein below.

BRIEF DESCRIPTION OF THE FIGURE

Having thus described the presently disclosed subject matter in generalterms, reference will now be made to the accompanying FIGURE, which isnot necessarily drawn to scale, and wherein:

FIG. 1 shows whole body and ex vivo organ imaging of mouse with PSMA⁺PC3 PIP tumor and PSMA⁻ PC3 flu tumor at 24 h postinjection of 1 nmol ofDyLight800-3.

The patent or application file contains at least one drawing executed incolor. Copies of this patent or patent application publication withcolor drawings will be provided by the Office upon request and paymentof the necessary fee.

DETAILED DESCRIPTION

The presently disclosed subject matter now will be described more fullyhereinafter with reference to the accompanying Figures, in which some,but not all embodiments of the inventions are shown. Like numbers referto like elements throughout. The presently disclosed subject matter maybe embodied in many different forms and should not be construed aslimited to the embodiments set forth herein; rather, these embodimentsare provided so that this disclosure will satisfy applicable legalrequirements. Indeed, many modifications and other embodiments of thepresently disclosed subject matter set forth herein will come to mind toone skilled in the art to which the presently disclosed subject matterpertains having the benefit of the teachings presented in the foregoingdescriptions and the associated Figures. Therefore, it is to beunderstood that the presently disclosed subject matter is not to belimited to the specific embodiments disclosed and that modifications andother embodiments are intended to be included within the scope of theappended claims.

I. PSMA Targeted Fluorescent Agents for Image Guided Surgery

Minimally invasive medical techniques are intended to reduce the amountof extraneous tissue that is damaged during diagnostic or surgicalprocedures, thereby reducing patient recovery time, discomfort, anddeleterious side effects. While millions of “open” or traditionalsurgeries are performed each year in the United States; many of thesesurgeries potentially can be performed in a minimally invasive manner.One effect of minimally invasive surgery, for example, is reducedpost-operative recovery time and related hospital stay. Because theaverage hospital stay for a standard surgery is typically significantlylonger than the average stay for an analogous minimally invasivesurgery, increased use of minimally invasive techniques could savemillions of dollars in hospital costs each year. While many of thesurgeries performed in the United States could potentially be performedin a minimally invasive manner, only a portion currently employ thesetechniques due to instrument limitations, method limitations, and theadditional surgical training involved in mastering the techniques.

Minimally invasive telesurgical systems are being developed to increasea surgeon's dexterity, as well as to allow a surgeon to operate on apatient from a remote location. Telesurgery is a general term forsurgical systems where the surgeon uses some form of remote control,e.g., a servomechanism, or the like, to manipulate surgical instrumentmovements rather than directly holding and moving the instruments byhand. In such a telesurgery system, the surgeon is provided with animage of the surgical site at the remote location. While viewing thesurgical site on a suitable viewer or display, the surgeon performs thesurgical procedures on the patient by manipulating master control inputdevices, which in turn control the motion of instruments. These inputdevices can move the working ends of the surgical instruments withsufficient dexterity to perform quite intricate surgical tasks.

Surgery is the most commonly used treatment for clinically localizedprostate cancer (PCa) and provides a survival advantage compared towatchful waiting. A pressing issue in surgery for PCa is the assuranceof a complete resection of the tumor, namely, a negative surgicalmargin. Surgical techniques, including minimally invasive surgicaltechniques, such as tele-surgical systems, can be further aided byimproving visualization of the tissue where the procedure is to becarried out. One way to improve visualization of tissue is through theuse of dyes capable of targeted visualization of tissue, allowing asurgeon to either remove or spare the tissue.

Accordingly, in some embodiments, the presently disclosed subject matterprovides low-molecular-weight compounds comprising PSMA-targetingligands linked to near-infrared (NIR), closed chain, sulfo-cyanine dyesand methods of their use for visualizing tissue under illumination withNIR radiation, including methods for imaging prostate cancer (PCa).

While a variety of radiolabeled PSMA-targeting antibodies have been usedfor tumor imaging, low molecular weight agents are preferred due to moretractable pharmacokinetics, i.e., more rapid clearance from nontargetsites. A series of fluorescent agents has been previously reported andwas tested in mice to good effect. See, for example, international PCTpatent application publication no. WO2010/108125A2, for PSMA-TARGETINGCOMPOUNDS AND USES THEREOF, to Pomper et al., published Sep. 23, 2010,which is incorporated by reference in its entirety. Because of thefavorable pharmacokinetic profile of this class of compounds, i.e., lownonspecific binding, lack of metabolism in vivo and reasonable tumorresidence times, this series of compounds was extended to includeDylight800 fluorescent dyes. Thus, the presently disclosed compoundsinclude a urea-based PSMA binding moiety linked to a Dylight™ 800fluorescent dye (Thermo Fisher Scientific Inc., Rockford, Ill., USA).The presently disclosed targeted fluorescent PSMA binding compounds mayfind utility in fluorescence image guided surgery and biopsy of PSMApositive tumors and tissues; the former providing visual confirmation ofcomplete removal of PSMA-containing tissue.

A. Compound (3)

Accordingly, in some embodiments, the presently disclosed subject matterprovides the following compound:

or a pharmaceutically acceptable salt thereof.

The presently disclosed compounds can be made using procedures known inthe art by attaching near IR, closed chain, sulfo-cyanine dyes toprostate specific membrane antigen ligands via a linkage. For example,the prostate specific membrane antigen ligands used in the presentlydisclosed compounds can be synthesized as described in international PCTpatent application publication no. WO 2010/108125, to Pomper et al.,published Sep. 23, 2010, which is incorporated herein in its entirety.Compounds can assembled by reactions between different components, toform linkages such as ureas (—NRC(O)NR—), thioureas (—NRC(S)NR—), amides(—C(O)NR— or —NRC(O)—), or esters (—C(O)O— or —OC(O)—). Urea linkagescan be readily prepared by reaction between an amine and an isocyanate,or between an amine and an activated carbonamide (—NRC(O)—). Thioureascan be readily prepared from reaction of an amine with anisothiocyanate. Amides (—C(O)NR— or —NRC(O)—) can be readily prepared byreactions between amines and activated carboxylic acids or esters, suchas an acyl halide or N-hydroxysuccinimide ester. Carboxylic acids mayalso be activated in situ, for example, with a coupling reagent, such asa carbodiimide, or carbonyldiimidazole (CDI). Esters may be formed byreaction between alcohols and activated carboxylic acids. Triazoles arereadily prepared by reaction between an azide and an alkyne, optionallyin the presence of a copper (Cu) catalyst.

Prostate specific membrane antigen ligands can also be prepared bysequentially adding components to a preformed urea, such as thelysine-urea-glutamate compounds described in Banerjee et al. (J. Med.Chem. vol. 51, pp. 4504-4517, 2008). Other urea-based compounds may alsobe used as building blocks.

Exemplary syntheses of the near IR, closed chain, sulfo-cyanine dyesused in the presently disclosed compositions are described in U.S. Pat.No. 6,887,854 and U.S. Pat. No. 6,159,657 and are incorporated herein intheir entirety. Additionally, some IR, closed chain, sulfo-cyanine dyesof the presently disclosed subject matter are commercially available,including DyLight™ 800 (ThermoFisher).

As provided hereinabove, the presently disclosed compounds can besynthesized via attachment of near IR, closed chain, sulfo-cyanine dyesto prostate specific membrane antigen ligands by reacting a reactiveamine on the ligand with a near IR dye. A wide variety of near IR dyesare known in the art, with activated functional groups for reacting withamines.

B. Compositions Comprising Compound (3)

In some embodiments, the presently disclosed subject matter provides acomposition comprising a unit dosage form of compound (3), or apharmaceutically acceptable salt thereof, wherein the composition isadapted for administration to a subject; and wherein, the unit dosageform delivers to the subject an amount between 0.01 mg/kg and 8 mg/kg ofcompound (3). In some embodiments, the composition unit dosage formdelivers to the subject the amount of about 0.01 mg/kg, about 0.05mg/kg, about 0.10 mg/kg, about 0.20 mg/kg, about 0.30 mg/kg, about 0.35mg/kg, about 0.40 mg/kg, about 0.45 mg/kg, about 0.50 mg/kg, about 0.55mg/kg, about 0.60 mg/kg, about 0.65 mg/kg, about 0.70 mg/kg, about 0.75mg/kg, about 0.80 mg/kg, about 0.90 mg/kg, about 1 mg/kg, about 2 mg/kg,about 4 mg/kg, about 6 mg/kg, or about 8 mg/kg. In some embodiments, thecomposition is dry and a single dose form.

The term “unit dosage form” as used herein encompasses any measuredamount that can suitably be used for administering a pharmaceuticalcomposition to a patient. As recognized by those skilled in the art,when another form (e.g., another salt the pharmaceutical composition) isused in the formulation, the weight can be adjusted to provide anequivalent amount of the pharmaceutical composition.

In some embodiments, the composition is lyophilized in a sterilecontainer. In some embodiments, the composition is contained within asterile container, wherein the container has a machine detectableidentifier that is readable by a medical device.

As used herein, the term “sterile” refers to a system or components of asystem free of infectious agents, including, but not limited to,bacteria, viruses, and bioactive RNA or DNA.

As used herein, the term “non-toxic” refers to the non-occurrence ofdetrimental effects when administered to a vertebrate as a result ofusing a pharmaceutical composition at levels effective for visualizationof tissue under illumination with near-infrared radiation (therapeuticlevels).

As used herein, the term “machine detectable identifier” includesidentifiers visible or detectable by machines including medical devices.In some instances, the medical device is a telesurgical system. Machinedetectable identifiers may facilitate the access or utilization ofinformation that is directly encoded in the machine detectableidentifier, or stored elsewhere. Examples of machine detectibleidentifiers include, but are not limited to, microchips, radio frequencyidentification (RFID) tags, barcodes (e.g., 1-dimensional or2-dimensional barcode), data matrices, quick-response (QR) codes, andholograms. One of skill in the art will recognize that other machinedetectible identifiers are useful in the presently disclosed subjectmatter.

In some embodiments, the composition further comprises compound (3) incombination with pharmaceutically acceptable excipients in an oraldosage form. In some embodiments, the composition further comprisescompound (3) in combination with pharmaceutically acceptable carriers inan injectable dosage form. In some embodiments, the composition furthercomprises compound (3) in combination with pharmaceutically acceptableexcipients in a dosage form for direct delivery to a surgical site.

“Pharmaceutically acceptable excipient” and “pharmaceutically acceptablecarrier” refer to a substance that aids the administration of an activeagent to and absorption by a patient and can be included in thepresently disclosed compositions without causing a significant adversetoxicological effect on the patient. Non-limiting examples ofpharmaceutically acceptable excipients include water, NaCl, normalsaline solutions, lactated Ringer's, normal sucrose, normal glucose,binders, fillers, disintegrants, lubricants, coatings, sweeteners,flavors and colors, and the like. One of skill in the art will recognizethat other pharmaceutical excipients are useful in the presentlydisclosed subject matter. Pharmaceutically acceptable carriers includebut not limited to any adjuvants, excipients, glidants, sweeteners,diluents, preservatives, dyes/colorants, flavoring agents, surfactants,wetting agents, dispersing agents, suspending agents, stabilizingagents, isotonic agents, solvents or emulsors.

The term “oral dosage form” as used herein refers to its normal meaningin the art (i.e., a pharmaceutical composition in the form of a tablet,capsule, caplet, gelcap, geltab, pill and the like).

The term “injectable dosage form” as used herein refers to its normalmeaning in the art (i.e., refer to a pharmaceutical composition in theform of solutions, suspensions, and emulsions, for example, water orwater/propylene glycol solutions.)

The presently disclosed compositions can be prepared in a wide varietyof oral, parenteral and topical dosage forms. Oral preparations includetablets, pills, powder, dragees, capsules, liquids, lozenges, cachets,gels, syrups, slurries, suspensions, etc., suitable for ingestion by thepatient. The presently disclosed compositions can also be administeredby injection, that is, intravenously, intramuscularly, intracutaneously,subcutaneously, intraduodenally, or intraperitoneally. Also, thecompositions described herein can be administered by inhalation, forexample, intranasally. Additionally, the presently disclosedcompositions can be administered transdermally. The compositions of thisinvention can also be administered by intraocular, insufflation,powders, and aerosol formulations (for examples of steroid inhalants,see Rohatagi, J. Clin. Pharmacol. 35:1187-1193, 1995; Tjwa, Ann. AllergyAsthma Immunol. 75:107-111, 1995). Accordingly, the presently disclosedsubject matter also provides pharmaceutical compositions including apharmaceutically acceptable carrier or excipient.

For preparing pharmaceutical compositions from the presently disclosedsubject matter, pharmaceutically acceptable carriers can be either solidor liquid. Solid form preparations include powders, tablets, pills,capsules, cachets, suppositories, and dispersible granules. A solidcarrier can be one or more substances, which may also act as diluents,flavoring agents, binders, preservatives, tablet disintegrating agents,or an encapsulating material. Details on techniques for formulation andadministration are well described in the scientific and patentliterature, see, e.g., the latest edition of Remington's PharmaceuticalSciences, Maack Publishing Co, Easton Pa. (“Remington's”).

In powders, the carrier is a finely divided solid, which is in a mixturewith the finely divided active component. In tablets, the activecomponent is mixed with the carrier having the necessary bindingproperties in suitable proportions and compacted in the shape and sizedesired. The powders and tablets preferably contain from 5% or 10% to70% of the compounds of the presently disclosed subject matter.

Suitable solid excipients include, but are not limited to, magnesiumcarbonate; magnesium stearate; talc; pectin; dextrin; starch;tragacanth; a low melting wax; cocoa butter; carbohydrates; sugarsincluding, but not limited to, lactose, sucrose, mannitol, or sorbitol,starch from corn, wheat, rice, potato, or other plants; cellulose suchas methyl cellulose, hydroxypropylmethyl-cellulose, or sodiumcarboxymethylcellulose; and gums including arabic and tragacanth; aswell as proteins including, but not limited to, gelatin and collagen. Ifdesired, disintegrating or solubilizing agents may be added, such as thecross-linked polyvinyl pyrrolidone, agar, alginic acid, or a saltthereof, such as sodium alginate.

Dragee cores are provided with suitable coatings such as concentratedsugar solutions, which may also contain gum arabic, talc,polyvinylpyrrolidone, carbopol gel, polyethylene glycol, and/or titaniumdioxide, lacquer solutions, and suitable organic solvents or solventmixtures. Dyestuffs or pigments may be added to the tablets or drageecoatings for product identification or to characterize the quantity ofactive compound (i.e., dosage). Pharmaceutical compositions of theinvention can also be used orally using, for example, push-fit capsulesmade of gelatin, as well as soft, sealed capsules made of gelatin and acoating such as glycerol or sorbitol. Push-fit capsules can contain thepresently disclosed compositions mixed with a filler or binders such aslactose or starches, lubricants such as talc or magnesium stearate, and,optionally, stabilizers. In soft capsules, the presently disclosedcompositions may be dissolved or suspended in suitable liquids, such asfatty oils, liquid paraffin, or liquid polyethylene glycol with orwithout stabilizers.

Liquid form preparations include solutions, suspensions, and emulsions,for example, water or water/propylene glycol solutions. For parenteralinjection, liquid preparations can be formulated in solution in aqueouspolyethylene glycol solution.

Aqueous solutions suitable for oral use can be prepared by dissolvingthe presently disclosed compositions in water and adding suitablecolorants, flavors, stabilizers, and thickening agents as desired.Aqueous suspensions suitable for oral use can be made by dispersing thefinely divided active component in water with viscous material, such asnatural or synthetic gums, resins, methylcellulose, sodiumcarboxymethylcellulose, hydroxypropylmethylcellulose, sodium alginate,polyvinylpyrrolidone, gum tragacanth and gum acacia, and dispersing orwetting agents such as a naturally occurring phosphatide (e.g.,lecithin), a condensation product of an alkylene oxide with a fatty acid(e.g., polyoxyethylene stearate), a condensation product of ethyleneoxide with a long chain aliphatic alcohol (e.g., heptadecaethyleneoxycetanol), a condensation product of ethylene oxide with a partialester derived from a fatty acid and a hexitol (e.g., polyoxyethylenesorbitol mono-oleate), or a condensation product of ethylene oxide witha partial ester derived from fatty acid and a hexitol anhydride (e.g.,polyoxyethylene sorbitan mono-oleate). The aqueous suspension can alsocontain one or more preservatives such as ethyl or n-propylp-hydroxybenzoate, one or more coloring agents, one or more flavoringagents and one or more sweetening agents, such as sucrose, aspartame orsaccharin. Formulations can be adjusted for osmolarity.

Also included are solid form preparations, which are intended to beconverted, shortly before use, to liquid form preparations for oraladministration. Such liquid forms include solutions, suspensions, andemulsions. These preparations may contain, in addition to the activecomponent, colorants, flavors, stabilizers, buffers, artificial andnatural sweeteners, dispersants, thickeners, solubilizing agents, andthe like.

Oil suspensions can be formulated by suspending the presently disclosedcompositions in a vegetable oil, such as arachis oil, olive oil, sesameoil or coconut oil, or in a mineral oil such as liquid paraffin; or amixture of these. The oil suspensions can contain a thickening agent,such as beeswax, hard paraffin or cetyl alcohol. Sweetening agents canbe added to provide a palatable oral preparation, such as glycerol,sorbitol or sucrose. These formulations can be preserved by the additionof an antioxidant such as ascorbic acid. As an example of an injectableoil vehicle, see Minto, J. Pharmacol. Exp. Ther. 281:93-102, 1997. Thepharmaceutical formulations of the invention can also be in the form ofoil-in-water emulsions. The oily phase can be a vegetable oil or amineral oil, described above, or a mixture of these. Suitableemulsifying agents include naturally-occurring gums, such as gum acaciaand gum tragacanth, naturally occurring phosphatides, such as soybeanlecithin, esters or partial esters derived from fatty acids and hexitolanhydrides, such as sorbitan mono-oleate, and condensation products ofthese partial esters with ethylene oxide, such as polyoxyethylenesorbitan mono-oleate. The emulsion can also contain sweetening agentsand flavoring agents, as in the formulation of syrups and elixirs. Suchformulations can also contain a demulcent, a preservative, or a coloringagent.

The presently disclosed compositions can also be delivered asmicrospheres for slow release in the body. For example, microspheres canbe formulated for administration via intradermal injection ofdrug-containing microspheres, which slowly release subcutaneously (seeRao, J. Biomater Sci. Polym. Ed. 7:623-645, 1995; as biodegradable andinjectable gel formulations (see, e.g., Gao Pharm. Res. 12:857-863,1995); or, as microspheres for oral administration (see, e.g., Eyles, J.Pharm. Pharmacol. 49:669-674, 1997). Both transdermal and intradermalroutes afford constant delivery for weeks or months.

In another embodiment, the presently disclosed compositions can beformulated for parenteral administration, such as intravenous (IV)administration or administration into a body cavity or lumen of anorgan. The formulations for administration will commonly comprise asolution of the presently disclosed compositions dissolved in apharmaceutically acceptable carrier. Among the acceptable vehicles andsolvents that can be employed are water and Ringer's solution, anisotonic sodium chloride. In addition, sterile fixed oils canconventionally be employed as a solvent or suspending medium. For thispurpose any bland fixed oil can be employed including synthetic mono- ordiglycerides. In addition, fatty acids such as oleic acid can likewisebe used in the preparation of injectables. These solutions are sterileand generally free of undesirable matter. These formulations may besterilized by conventional, well-known techniques including radiation,chemical, heat/pressure, and filtration sterilization techniques. Theformulations may contain pharmaceutically acceptable auxiliarysubstances as required to approximate physiological conditions such aspH adjusting and buffering agents, toxicity adjusting agents, e.g.,sodium acetate, sodium chloride, potassium chloride, calcium chloride,sodium lactate and the like. The concentration of the presentlydisclosed compositions in these formulations can vary widely, and willbe selected primarily based on fluid volumes, viscosities, body weight,and the like, in accordance with the particular mode of administrationselected and the patient's needs. For IV administration, the formulationcan be a sterile injectable preparation, such as a sterile injectableaqueous or oleaginous suspension. This suspension can be formulatedaccording to the known art using those suitable dispersing or wettingagents and suspending agents. The sterile injectable preparation canalso be a sterile injectable solution or suspension in a non-toxicparenterally-acceptable diluent or solvent, such as a solution of1,3-butanediol.

In another embodiment, the formulations of the presently disclosedcompositions can be delivered by the use of liposomes which fuse withthe cellular membrane or are endocytosed, i.e., by employing ligandsattached to the liposome, or attached directly to the oligonucleotide,that bind to surface membrane protein receptors of the cell resulting inendocytosis. By using liposomes, particularly where the liposome surfacecarries ligands specific for target cells, or are otherwisepreferentially directed to a specific organ, one can focus the deliveryof the presently disclosed compositions into the target cells in vivo.(See, e.g., Al-Muhammed, J. Microencapsul. 13:293-306, 1996; Chonn,Curr. Opin. Biotechnol. 6:698-708, 1995; Ostro, Am. J. Hosp. Pharm.46:1576-1587, 1989).

Lipid-based drug delivery systems include lipid solutions, lipidemulsions, lipid dispersions, self-emulsifying drug delivery systems(SEDDS) and self-microemulsifying drug delivery systems (SMEDDS). Inparticular, SEDDS and SMEDDS are isotropic mixtures of lipids,surfactants and co-surfactants that can disperse spontaneously inaqueous media and form fine emulsions (SEDDS) or microemulsions(SMEDDS). Lipids useful in the formulations of the presently disclosedsubject matter include any natural or synthetic lipids including, butnot limited to, sesame seed oil, olive oil, castor oil, peanut oil,fatty acid esters, glycerol esters, Labrafil®, Labrasol®, Cremophor®,Solutol®, Tween®, Capryol®, Capmul®, Captex®, and Peceol®.

In some embodiments the presently disclosed compositions are sterile andgenerally free of undesirable matter. The compounds and compositions maybe sterilized by conventional, well known techniques includingheat/pressure, gas plasma, steam, radiation, chemical, and filtrationsterilization techniques.

For example, terminal heat sterilization can be used to destroy allviable microorganisms within the final formulation. An autoclave iscommonly used to accomplish terminal heat-sterilization of drug productsin their final packaging. Typical autoclave cycles in the pharmaceuticalindustry to achieve terminal sterilization of the final product are 121°C. for 15 minutes. The presently disclosed compositions can beautoclaved at a temperature ranging from 115 to 130° C. for a period oftime ranging from 5 to 40 minutes with acceptable stability. Autoclavingis preferably carried out in the temperature range of 119 to 122° C. fora period of time ranging from 10 to 36 minutes.

The compositions can also be sterilized by dry heat as described byKarlsson, et al., in U.S. Pat. No. 6,392,036, which discloses a methodfor the dry heat sterilization that can be used for drug formulations.The compositions can also be sterilized as described in WO 02/41925 toBreath Limited, which discloses a rapid method, similar topasteurization, for the sterilization of compositions. This methodentails pumping the composition to be sterilized through stainless steelpipes and rapidly raising the temperature of the composition to about130-145° C. for about 2-20 seconds, subsequently followed by rapidcooling in seconds to ambient conditions.

The compositions can also be sterilized by irradiation as described byIllum and Moeller in Arch. Pharm. Chem. Sci., Ed. 2, 1974, pp. 167-174).The compositions can also be sterilized by UV, x-rays, gamma rays, ebeam radiation, flaming, baking, and chemical sterilization.

Alternatively, sterile pharmaceutical compositions according to thepresently disclosed subject matter may be prepared using asepticprocessing techniques. Aseptic filling is ordinarily used to preparedrug products that will not withstand heat sterilization, but in whichall of the ingredients are sterile. Sterility is maintained by usingsterile materials and a controlled working environment. All containersand apparatus are sterilized, preferably by heat sterilization, prior tofilling. The container (e.g., vial, ampoule, infusion bag, bottle, orsyringe) are then filled under aseptic conditions.

In some embodiments, the compounds and presently disclosed compositionsare non-toxic and generally free of detrimental effects whenadministered to a vertebrate at levels effective for visualization oftissue under illumination with near-infrared radiation. Toxicity of thecompounds and presently disclosed compositions can be assessed bymeasuring their effects on a target (organism, organ, tissue or cell).Because individual targets typically have different levels of responseto the same dose of a compound, a population-level measure of toxicityis often used which relates the probabilities of an outcome for a givenindividual in a population. Toxicology of compounds can be determined byconventional, well-known techniques including in vitro (outside of aliving organism) and in vivo (inside of a living organism) studies.

For example, determination of metabolic stability is commonly examinedwhen assessing the toxicity of a compound as it is one of several majordeterminates in defining the oral bioavailability and systemic clearanceof a compound. After a compound is administered orally, it firstencounters metabolic enzymes in the gastrointestinal lumen as well as inthe intestinal epithelium. After it is absorbed into the bloodstreamthrough the intestinal epithelium, it is first delivered to the livervia the portal vein. A compound can be effectively cleared by intestinalor hepatic metabolism before it reaches systemic circulation, a processknown as first pass metabolism. The stability of a compound towardsmetabolism within the liver as well as extrahepatic tissues willultimately determine the concentration of the compound found in thesystemic circulation and affect its half-life and residence time withinthe body. Cytochrome P450 (CYP) enzymes are found primarily in the liverbut also in the intestinal wall, lungs, kidneys and other extrahepaticorgans and are the major enzymes involved in compound metabolism. Manycompounds undergo deactivation by CYPs, either directly or byfacilitated excretion from the body. Also, many compounds arebioactivated by CYPs to form their active compounds. Thus, determiningthe reactivity of a compound to CYP enzymes is commonly used to assessmetabolic stability of a compound.

The Ames reverse mutation Assay is another common toxicology assay forassessing the toxicity of a compound. The Ames Assay, utilizes severaldifferent tester strains, each with a distinct mutation in one of thegenes comprising the histidine (his) biosynthetic operon (Ames, B. N.,et al., (1975) Mutation Res. 31:347-363). The detection of revertant(i.e., mutant) bacteria in test samples that are capable of growth inthe absence of histidine indicates that the compound under evaluation ischaracterized by genotoxic (i.e. mutagenic) activity. The Ames Assay iscapable of detecting several different types of mutations (geneticdamage) that may occur in one or more of the tester strains. Thepractice of using an in vitro bacterial assay to evaluate the genotoxicactivity of drug candidates is based on the prediction that a substancethat is mutagenic in a bacterium is likely to be carcinogenic inlaboratory animals, and by extension may be carcinogenic or mutagenic tohumans.

In addition, the human ether-a-go-go related gene (hERG) assay can beused to evaluate the potential cardiotoxicity of a compound.Cardiotoxicity can arise when the QT interval is prolonged leading to anelevated risk of life-threatening arrhythmias. The QT interval is ameasure of the time between the start of the Q wave and the end of the Twave in the heart's electrical cycle. The QT interval representselectrical depolarization and repolarization of the ventricles. Alengthened QT interval has most commonly been associated with loss ofcurrent through hERG potassium ion channels due to direct block of theion channel by drugs or by inhibition of the plasma membrane expressionof the channel protein (Su et al. J. Biomol Screen 2011, 16, 101-111).Thus, an in vitro hERG screening assay can be used to detect disruptionor inhibition of the hERG membrane trafficking function and assesspotential cardiotoxicity of a compound.

Other methods of assessing the toxicity of compounds include in vivostudies which administer relatively large doses of a test compound to agroup of animals to determine the level which is lethal to a percentageof the population (mean lethal dose LD₅₀ or mean lethal concentrationLC₅₀). Toxicity of a compound can also be assessed in vivo by examiningwhether a compound produces statistically significant negative effectson cardiac, blood pressure, central nervous system (CNS), body weight,food intake, gross or microscopic pathology, clinical pathology, orrespiratory measures in an animal.

In some embodiments, the presently disclosed compositions can belyophilized in a sterile container for convenient dry storage andtransport. A ready-to-use preparation can subsequently be made byreconstituting the lyophilized compositions with sterile water. Theterms “lyophilization,” “lyophilized,” and “freeze-dried” refer to aprocess by which the material to be dried is first frozen and then theice or frozen solvent is removed by sublimation in a vacuum environment.An excipient may be included in pre-lyophilized formulations to enhancestability of the lyophilized product upon storage.

In some embodiments, the composition can be contained within a sterilecontainer, where the container has a machine detectable identifier whichis readable by a medical device. Examples of machine detectibleidentifiers include microchips, radio frequency identification (RFID)tags, barcodes (e.g., 1-dimensional or 2-dimensional barcode), datamatrices, quick-response (QR) codes, and holograms. One of skill in theart will recognize that other machine detectible identifiers are usefulin the presently disclosed subject matter.

In some embodiments, the machine detectable identifier can include amicrochip, an integrated circuit (IC) chip, or an electronic signal froma microchip that is detectable and/or readable by a computer system thatis in communication with the medical device. In some embodiments, themachine detectable identifier includes a radio frequency identification(RFID) tag. RFID tags are sometimes called as transponders. RFID tagsgenerally are devices formed of an IC chip, an antenna, an adhesivematerial, and are used for transmitting or receiving predetermined datawith an external reader or interrogator. RFID tags can transmit orreceive data with a reader by using a contactless method. According tothe amplitude of a used frequency, inductive coupling, backscattering,and surface acoustic wave (SAW) may be used. Using electromagneticwaves, data may be transmitted or received to or from a reader by usinga full duplex method, a half duplex (HDX) method, or a sequential (SEQ)method.

In some embodiments, the machine detectable identifier can include abarcode. Barcodes include any machine-readable format, includingone-dimensional and two-dimensional formats. One-dimensional formatsinclude, for example, Universal Product Code (UPC) and Reduced SpaceSymbology (RSS). Two-dimensional formats, or machine-readable matrices,include for example, Quick Response (QR) Code and Data Matrix.

In some embodiments, the medical device can be configured to detect themachine detectable identifier. In one example, the medical device is atele-surgical system that includes a special imaging mode (e.g., afluorescence imaging mode) for use with dyes such as those described inthis disclosure. One example of a tele-surgical system that includes afluorescence imaging mode is described in U.S. Pat. No. 8,169,468,entitled “Augmented Stereoscopic Visualization for a Surgical Robot,”which is hereby incorporated in its entirety herein. In someembodiments, medical devices can incorporate an imaging device that canscan, read, view, or otherwise detect a machine detectable identifierthat is displayed to the imaging device. In one aspect, the medicaldevice will permit a user to access the fluorescence imaging mode of themedical device only if the medical device detects the presence of aknown machine detectable identifier that corresponds to a dye identifiedas being compatible for use with the medical device. In contrast, if themedical device does not detect a known machine detectable identifier,the medical device will not permit a user to access the fluorescenceimaging mode and associated functionality. Imaging devices can includeoptical scanners, barcode readers, cameras, and imaging devicescontained within a tele-surgical system such as an endoscope.Information associated with the machine detectable identifier can thenbe retrieved by the medical device using an imaging device. Upondetection of the identifier, an automatic process may be launched tocause a predetermined action to occur, or certain data to be retrievedor accessed. The information encoded onto the machine detectableidentifier may include instructions for triggering an action, such asadministering a composition of the presently disclosed subject matter toa patient. In some embodiments, the machine detectable identifierincludes unencrypted e-pedigree information in the desired format. Thee-pedigree information can include, for example, lot, potency,expiration, national drug code, electronic product code, manufacturer,distributor, wholesaler, pharmacy and/or a unique identifier of thesalable unit.

In some embodiments, the sterile container having a machine detectableidentifier includes a fluid outlet configured to mate with the medicaldevice. In some embodiments, the fluid outlet of the machine detectableidentifier is mechanically affixed to the medical device.

C. Methods of Imaging Using Compositions Comprising Compound (3)

In some embodiments, the presently disclosed subject matter provides ause of the composition comprising compound (3), or a pharmaceuticallyacceptable salt thereof, adapted for administration to a subject, e.g.,a patient, to obtain visualization of tissue expressing PSMA underillumination with near-infrared radiation, wherein the unit dosage formdelivers to the subject an amount between about 0.01 mg/kg and 8 mg/kgof compound (3). In some embodiments, the use is adapted foradministration to a human patient to obtain visualization of humantissue under illumination with near-infrared radiation wherein the unitdosage form delivers to the human patient an amount between about 0.01mg/kg and 8 mg/kg of a compound (3).

The compounds and presently disclosed compositions can be delivered byany suitable means, including oral, parenteral and topical methods.Transdermal administration methods, by a topical route, can beformulated as applicator sticks, solutions, suspensions, emulsions,gels, creams, ointments, pastes, jellies, paints, powders, and aerosol.

The pharmaceutical preparation is preferably in unit dosage form. Insuch form the preparation is subdivided into unit doses containingappropriate quantities of the compounds and presently disclosedcompositions. The unit dosage form can be a packaged preparation, thepackage containing discrete quantities of preparation, such as packetedtablets, capsules, and powders in vials or ampoules. Also, the unitdosage form can be a capsule, tablet, cachet, or lozenge itself, or itcan be the appropriate number of any of these in packaged form.

In some embodiments, co-administration can be accomplished byco-formulation, i.e., preparing a single pharmaceutical compositionincluding the compounds and presently disclosed compositions and anyother agent. Alternatively, the various components can be formulatedseparately.

The presently disclosed compositions, and any other agents, can bepresent in any suitable amount, and can depend on various factorsincluding, but not limited to, weight and age of the patient, state ofthe disease, etc. Suitable dosage ranges include from about 0.01 and 8mg/kg, or about 0.01 and 5 mg/kg, or about 0.01 and 1 mg/kg. Suitabledosage ranges also include 0.01, 0.05, 0.10, 0.20, 0.30, 0.35, 0.40,0.45, 0.50, 0.55, 0.60, 0.65, 0.70, 0.75, 0.80, 0.90, 1, 2, 4, 6, or 8mg/kg.

The composition can also contain other compatible compositions. Thecompositions described herein can be used in combination with oneanother, with other active compositions known to be useful forvisualization of tissue under illumination with near-infrared radiation,or with compositions that may not be effective alone, but may contributeto the efficacy of the active composition.

As used herein, the term “visualization” refers to methods of obtaininggraphic images of tissue by any means, including illumination withnear-infrared radiation.

The term “near-infrared radiation” or “near IR radiation” or “NIR”radiation refers to optical radiation with a wavelength in the range ofabout 700 nm to about 1400 nm. References herein to the optionallyplural term “wavelength(s)” indicates that the radiation may be a singlewavelength or a spectrum of radiation having differing wavelengths.

The term “tissue” as used herein includes, but is not limited to,allogenic or xenogenic bone, neural tissue, fibrous connective tissueincluding tendons and ligaments, cartilage, dura, fascia, pericardia,muscle, heart valves, veins and arteries and other vessels, dermis,adipose tissue, glandular tissue, prostate tissue, kidney tissue, braintissue, renal tissue, bladder tissue, lung tissue, breast tissue,pancreatic tissue, vascular tissue, tumor tissue, cancerous tissue, orprostate tumor tissue.

In particular embodiments, the presently disclosed subject matterprovides a method for visualization of tissue expressing PSMA, themethod comprising, administering to a subject, e.g., a patient, acomposition comprising compound (3), described herein. In someembodiments, the method comprises, administering to a subject acomposition comprising compound (3):

or a pharmaceutically acceptable salt thereof.

In some embodiments, the method administers to a subject apharmaceutical composition comprising a unit dosage form of compound(3), wherein the composition is sterile, non-toxic, and adapted foradministration to a subject; and wherein, the unit dosage form deliversto the subject an amount between about 0.01 mg/kg and 8 mg/kg ofcompound (3). In some embodiments, the method further comprisesobtaining the image during administration, after administration, or bothduring and after administration of the composition. In some embodiments,the method further comprises intravenously injecting a compositioncomprising compound (3) into a subject. In some embodiments, thecomposition is injected into a circulatory system of the subject.

In some embodiments, the method further comprises visualizing a subjectarea on which surgery is or will be performed, or for viewing a subjectarea otherwise being examined by a medical professional. In someembodiments, the method further comprises performing a surgicalprocedure on the subject areas based on the visualization of thesurgical area. In some embodiments, the method further comprises viewinga subject area on which an ophthalmic, arthroscopic, laparoscopic,cardiothoracic, muscular, or neurological procedure is or will beperformed. In some embodiments, the method further comprises obtainingex vivo images of at least a portion of the subject.

In some embodiments, the tissue being visualized is tumor tissue. Insome embodiments, the tissue being visualized is dysplastic or canceroustissue. In some embodiments, the tissue being visualized is prostatetissue. In some embodiments, the tissue being visualized is prostatetumor tissue.

In other embodiments, the one or more PSMA-expressing tumor or cell isselected from the group consisting of: a prostate tumor or cell, ametastasized prostate tumor or cell, a lung tumor or cell, a renal tumoror cell, a glioblastoma, a pancreatic tumor or cell, a bladder tumor orcell, a sarcoma, a melanoma, a breast tumor or cell, a colon tumor orcell, a germ cell, a pheochromocytoma, an esophageal tumor or cell, astomach tumor or cell, and combinations thereof. In more particularembodiments, the one or more PSMA-expressing tumor or cell is a prostatetumor or cell. In certain embodiments, the one or more PSMA-expressingtumors or cells are in vitro, in vivo, or ex vivo. In particularembodiments, the one or more PSMA-expressing tumors or cells are presentin a subject.

The “subject” treated by the presently disclosed methods in their manyembodiments is desirably a human subject, although it is to beunderstood that the methods described herein are effective with respectto all vertebrate species, which are intended to be included in the term“subject.” Accordingly, a “subject” can include a human subject formedical purposes, such as for the treatment of an existing condition ordisease or the prophylactic treatment for preventing the onset of acondition or disease, or an animal subject for medical, veterinarypurposes, or developmental purposes. Suitable animal subjects includemammals including, but not limited to, primates, e.g., humans, monkeys,apes, and the like; bovines, e.g., cattle, oxen, and the like; ovines,e.g., sheep and the like; caprines, e.g., goats and the like; porcines,e.g., pigs, hogs, and the like; equines, e.g., horses, donkeys, zebras,and the like; felines, including wild and domestic cats; canines,including dogs; lagomorphs, including rabbits, hares, and the like; androdents, including mice, rats, and the like. An animal may be atransgenic animal. In some embodiments, the subject is a humanincluding, but not limited to, fetal, neonatal, infant, juvenile, andadult subjects. Further, a “subject” can include a patient afflictedwith or suspected of being afflicted with a condition or disease. Thus,the terms “subject” and “patient” are used interchangeably herein. Theterm “subject” also refers to an organism, tissue, cell, or collectionof cells from a subject.

In general, the “effective amount” of an active agent refers to theamount necessary to elicit the desired biological response. As will beappreciated by those of ordinary skill in this art, the effective amountof an agent or device may vary depending on such factors as the desiredbiological endpoint, the agent to be delivered, the makeup of thepharmaceutical composition, the target tissue, and the like.

In some embodiments, compound (3) is cleared from the subject's kidneysin about 24 hours.

In some embodiments, the presently disclosed methods use compounds thatare stable in vivo such that substantially all, e.g., more than about50%, 60%, 70%, 80%, or more preferably 90% of the injected compound isnot metabolized by the body prior to excretion. In other embodiments,the compound comprising the imaging agent is stable in vivo.

II. Definitions

Although specific terms are employed herein, they are used in a genericand descriptive sense only and not for purposes of limitation. Unlessotherwise defined, all technical and scientific terms used herein havethe same meaning as commonly understood by one of ordinary skill in theart to which this presently described subject matter belongs.

While the following terms in relation to the presently disclosedcompounds are believed to be well understood by one of ordinary skill inthe art, the following definitions are set forth to facilitateexplanation of the presently disclosed subject matter. These definitionsare intended to supplement and illustrate, not preclude, the definitionsthat would be apparent to one of ordinary skill in the art upon reviewof the present disclosure.

The terms substituted, whether preceded by the term “optionally” or not,and substituent, as used herein, refer to the ability, as appreciated byone skilled in this art, to change one functional group for anotherfunctional group on a molecule, provided that the valency of all atomsis maintained. When more than one position in any given structure may besubstituted with more than one substituent selected from a specifiedgroup, the substituent may be either the same or different at everyposition. The substituents also may be further substituted (e.g., anaryl group substituent may have another substituent off it, such asanother aryl group, which is further substituted at one or morepositions).

Where substituent groups or linking groups are specified by theirconventional chemical formulae, written from left to right, they equallyencompass the chemically identical substituents that would result fromwriting the structure from right to left, e.g., —CH₂O— is equivalent to—OCH₂—; —C(═O)O— is equivalent to —OC(═O)—; —OC(═O)NR— is equivalent to—NRC(═O)O—, and the like.

When the term “independently selected” is used, the substituents beingreferred to (e.g., R groups, such as groups R₁, R₂, and the like, orvariables, such as “m” and “n”), can be identical or different. Forexample, both R₁ and R₂ can be substituted alkyls, or R₁ can be hydrogenand R₂ can be a substituted alkyl, and the like.

The terms “a,” “an,” or “a(n),” when used in reference to a group ofsubstituents herein, mean at least one. For example, where a compound issubstituted with “an” alkyl or aryl, the compound is optionallysubstituted with at least one alkyl and/or at least one aryl. Moreover,where a moiety is substituted with an R substituent, the group may bereferred to as “R-substituted.” Where a moiety is R-substituted, themoiety is substituted with at least one R substituent and each Rsubstituent is optionally different.

A named “R” or group will generally have the structure that isrecognized in the art as corresponding to a group having that name,unless specified otherwise herein. For the purposes of illustration,certain representative “R” groups as set forth above are defined below.

Description of compounds of the present disclosure is limited byprinciples of chemical bonding known to those skilled in the art.Accordingly, where a group may be substituted by one or more of a numberof substituents, such substitutions are selected so as to comply withprinciples of chemical bonding and to give compounds which are notinherently unstable and/or would be known to one of ordinary skill inthe art as likely to be unstable under ambient conditions, such asaqueous, neutral, and several known physiological conditions. Forexample, a heterocycloalkyl or heteroaryl is attached to the remainderof the molecule via a ring heteroatom in compliance with principles ofchemical bonding known to those skilled in the art thereby avoidinginherently unstable compounds.

Unless otherwise explicitly defined, a “substituent group,” as usedherein, includes a functional group selected from one or more of thefollowing moieties, which are defined herein:

The term hydrocarbon, as used herein, refers to any chemical groupcomprising hydrogen and carbon. The hydrocarbon may be substituted orunsubstituted. As would be known to one skilled in this art, allvalencies must be satisfied in making any substitutions. The hydrocarbonmay be unsaturated, saturated, branched, unbranched, cyclic, polycyclic,or heterocyclic. Illustrative hydrocarbons are further defined hereinbelow and include, for example, methyl, ethyl, n-propyl, isopropyl,cyclopropyl, allyl, vinyl, n-butyl, tert-butyl, ethynyl, cyclohexyl, andthe like.

The term “alkyl,” by itself or as part of another substituent, means,unless otherwise stated, a straight (i.e., unbranched) or branchedchain, acyclic or cyclic hydrocarbon group, or combination thereof,which may be fully saturated, mono- or polyunsaturated and can includedi- and multivalent groups, having the number of carbon atoms designated(i.e., C₁-C₁₀ means one to ten carbons, including 1, 2, 3, 4, 5, 6, 7,8, 9, and 10 carbons). In particular embodiments, the term “alkyl”refers to C₁₋₂₀ inclusive, including 1, 2, 3, 4, 5, 6, 7, 8, 9, 10, 11,12, 13, 14, 15, 16, 17, 18, 19, and 20 carbons, linear (i.e.,“straight-chain”), branched, or cyclic, saturated or at least partiallyand in some embodiments fully unsaturated (i.e., alkenyl and alkynyl)hydrocarbon radicals derived from a hydrocarbon moiety containingbetween one and twenty carbon atoms by removal of a single hydrogenatom.

Representative saturated hydrocarbon groups include, but are not limitedto, methyl, ethyl, n-propyl, isopropyl, n-butyl, isobutyl, sec-butyl,tert-butyl, n-pentyl, sec-pentyl, isopentyl, neopentyl, n-hexyl,sec-hexyl, n-heptyl, n-octyl, n-decyl, n-undecyl, dodecyl, cyclohexyl,(cyclohexyl)methyl, cyclopropylmethyl, and homologs and isomers thereof.

“Branched” refers to an alkyl group in which a lower alkyl group, suchas methyl, ethyl or propyl, is attached to a linear alkyl chain. “Loweralkyl” refers to an alkyl group having 1 to about 8 carbon atoms (i.e.,a C₁₋₈ alkyl), e.g., 1, 2, 3, 4, 5, 6, 7, or 8 carbon atoms. “Higheralkyl” refers to an alkyl group having about 10 to about 20 carbonatoms, e.g., 10, 11, 12, 13, 14, 15, 16, 17, 18, 19, or 20 carbon atoms.In certain embodiments, “alkyl” refers, in particular, to C₁₋₈straight-chain alkyls. In other embodiments, “alkyl” refers, inparticular, to C₁₋₈ branched-chain alkyls.

Alkyl groups can optionally be substituted (a “substituted alkyl”) withone or more alkyl group substituents, which can be the same ordifferent. The term “alkyl group substituent” includes but is notlimited to alkyl, substituted alkyl, halo, acylamino, acyl, hydroxyl,aryloxyl, alkoxyl, alkylthio, arylthio, aralkyloxyl, aralkylthio,carboxyl, alkoxycarbonyl, oxo, and cycloalkyl. There can be optionallyinserted along the alkyl chain one or more oxygen, sulfur or substitutedor unsubstituted nitrogen atoms, wherein the nitrogen substituent ishydrogen, lower alkyl (also referred to herein as “alkylaminoalkyl”), oraryl.

Thus, as used herein, the term “substituted alkyl” includes alkylgroups, as defined herein, in which one or more atoms or functionalgroups of the alkyl group are replaced with another atom or functionalgroup, including for example, alkyl, substituted alkyl, halogen, aryl,substituted aryl, alkoxyl, hydroxyl, nitro, amino, alkylamino,dialkylamino, sulfate, and mercapto.

The term “heteroalkyl,” by itself or in combination with another term,means, unless otherwise stated, a stable straight or branched chain, orcyclic hydrocarbon group, or combinations thereof, consisting of atleast one carbon atoms and at least one heteroatom selected from thegroup consisting of O, N, P, Si and S, and wherein the nitrogen,phosphorus, and sulfur atoms may optionally be oxidized and the nitrogenheteroatom may optionally be quaternized. The heteroatom(s) 0, N, P andS and Si may be placed at any interior position of the heteroalkyl groupor at the position at which alkyl group is attached to the remainder ofthe molecule. Examples include, but are not limited to, —CH₂—CH₂—O—CH₃,—CH₂—CH₂—NH—CH₃, —CH₂—CH₂—N(CH₃)—CH₃, —CH₂—S—CH₂—CH₃,—CH₂—CH₂₅—S(O)—CH₃, —CH₂—CH₂—S(O)₂—CH₃, —CH═CH—O—CH₃, —Si(CH₃)₃,—CH₂—CH═N—OCH₃, —CH═CH—N(CH₃)— CH₃, O—CH₃, —O—CH₂—CH₃, and —CN. Up totwo or three heteroatoms may be consecutive, such as, for example,—CH₂—NH—OCH₃ and —CH₂—O—Si(CH₃)₃.

As described above, heteroalkyl groups, as used herein, include thosegroups that are attached to the remainder of the molecule through aheteroatom, such as —C(O)NR′, —NR′R″, —OR′, —SR, —S(O)R, and/or—S(O₂)R′. Where “heteroalkyl” is recited, followed by recitations ofspecific heteroalkyl groups, such as —NR′R or the like, it will beunderstood that the terms heteroalkyl and —NR′R″ are not redundant ormutually exclusive. Rather, the specific heteroalkyl groups are recitedto add clarity. Thus, the term “heteroalkyl” should not be interpretedherein as excluding specific heteroalkyl groups, such as —NR′R″ or thelike.

“Cyclic” and “cycloalkyl” refer to a non-aromatic mono- or multicyclicring system of about 3 to about 10 carbon atoms, e.g., 3, 4, 5, 6, 7, 8,9, or 10 carbon atoms. The cycloalkyl group can be optionally partiallyunsaturated. The cycloalkyl group also can be optionally substitutedwith an alkyl group substituent as defined herein, oxo, and/or alkylene.There can be optionally inserted along the cyclic alkyl chain one ormore oxygen, sulfur or substituted or unsubstituted nitrogen atoms,wherein the nitrogen substituent is hydrogen, unsubstituted alkyl,substituted alkyl, aryl, or substituted aryl, thus providing aheterocyclic group. Representative monocyclic cycloalkyl rings includecyclopentyl, cyclohexyl, and cycloheptyl. Multicyclic cycloalkyl ringsinclude adamantyl, octahydronaphthyl, decalin, camphor, camphane, andnoradamantyl, and fused ring systems, such as dihydro- andtetrahydronaphthalene, and the like.

The term “cycloalkylalkyl,” as used herein, refers to a cycloalkyl groupas defined hereinabove, which is attached to the parent molecular moietythrough an alkyl group, also as defined above. Examples ofcycloalkylalkyl groups include cyclopropylmethyl and cyclopentylethyl.

The terms “cycloheteroalkyl” or “heterocycloalkyl” refer to anon-aromatic ring system, unsaturated or partially unsaturated ringsystem, such as a 3- to 10-member substituted or unsubstitutedcycloalkyl ring system, including one or more heteroatoms, which can bethe same or different, and are selected from the group consisting ofnitrogen (N), oxygen (O), sulfur (S), phosphorus (P), and silicon (Si),and optionally can include one or more double bonds.

The cycloheteroalkyl ring can be optionally fused to or otherwiseattached to other cycloheteroalkyl rings and/or non-aromatic hydrocarbonrings. Heterocyclic rings include those having from one to threeheteroatoms independently selected from oxygen, sulfur, and nitrogen, inwhich the nitrogen and sulfur heteroatoms may optionally be oxidized andthe nitrogen heteroatom may optionally be quaternized. In certainembodiments, the term heterocylic refers to a non-aromatic 5-, 6-, or7-membered ring or a polycyclic group wherein at least one ring atom isa heteroatom selected from O, S, and N (wherein the nitrogen and sulfurheteroatoms may be optionally oxidized), including, but not limited to,a bi- or tri-cyclic group, comprising fused six-membered rings havingbetween one and three heteroatoms independently selected from theoxygen, sulfur, and nitrogen, wherein (i) each 5-membered ring has 0 to2 double bonds, each 6-membered ring has 0 to 2 double bonds, and each7-membered ring has 0 to 3 double bonds, (ii) the nitrogen and sulfurheteroatoms may be optionally oxidized, (iii) the nitrogen heteroatommay optionally be quaternized, and (iv) any of the above heterocyclicrings may be fused to an aryl or heteroaryl ring. Representativecycloheteroalkyl ring systems include, but are not limited topyrrolidinyl, pyrrolinyl, imidazolidinyl, imidazolinyl, pyrazolidinyl,pyrazolinyl, piperidyl, piperazinyl, indolinyl, quinuclidinyl,morpholinyl, thiomorpholinyl, thiadiazinanyl, tetrahydrofuranyl, and thelike.

The terms “cycloalkyl” and “heterocycloalkyl”, by themselves or incombination with other terms, represent, unless otherwise stated, cyclicversions of “alkyl” and “heteroalkyl”, respectively. Additionally, forheterocycloalkyl, a heteroatom can occupy the position at which theheterocycle is attached to the remainder of the molecule. Examples ofcycloalkyl include, but are not limited to, cyclopentyl, cyclohexyl,1-cyclohexenyl, 3-cyclohexenyl, cycloheptyl, and the like. Examples ofheterocycloalkyl include, but are not limited to,1-(1,2,5,6-tetrahydropyridyl), 1-piperidinyl, 2-piperidinyl,3-piperidinyl, 4-morpholinyl, 3-morpholinyl, tetrahydrofuran-2-yl,tetrahydrofuran-3-yl, tetrahydrothien-2-yl, tetrahydrothien-3-yl,1-piperazinyl, 2-piperazinyl, and the like. The terms “cycloalkylene”and “heterocycloalkylene” refer to the divalent derivatives ofcycloalkyl and heterocycloalkyl, respectively.

An unsaturated alkyl group is one having one or more double bonds ortriple bonds. Examples of unsaturated alkyl groups include, but are notlimited to, vinyl, 2-propenyl, crotyl, 2-isopentenyl, 2-(butadienyl),2,4-pentadienyl, 3-(1,4-pentadienyl), ethynyl, 1- and 3-propynyl,3-butynyl, and the higher homologs and isomers. Alkyl groups that arelimited to hydrocarbon groups are termed “homoalkyl.”

More particularly, the term “alkenyl” as used herein refers to amonovalent group derived from a C₁₋₂₀ inclusive straight or branchedhydrocarbon moiety having at least one carbon-carbon double bond by theremoval of a single hydrogen molecule. Alkenyl groups include, forexample, ethenyl (i.e., vinyl), propenyl, butenyl,1-methyl-2-buten-1-yl, pentenyl, hexenyl, octenyl, allenyl, andbutadienyl.

The term “cycloalkenyl” as used herein refers to a cyclic hydrocarboncontaining at least one carbon-carbon double bond. Examples ofcycloalkenyl groups include cyclopropenyl, cyclobutenyl, cyclopentenyl,cyclopentadiene, cyclohexenyl, 1,3-cyclohexadiene, cycloheptenyl,cycloheptatrienyl, and cyclooctenyl.

The term “alkynyl” as used herein refers to a monovalent group derivedfrom a straight or branched C₁₋₂₀ hydrocarbon of a designed number ofcarbon atoms containing at least one carbon-carbon triple bond. Examplesof “alkynyl” include ethynyl, 2-propynyl (propargyl), 1-propynyl,pentynyl, hexynyl, and heptynyl groups, and the like.

The term “alkylene” by itself or a part of another substituent refers toa straight or branched bivalent aliphatic hydrocarbon group derived froman alkyl group having from 1 to about 20 carbon atoms, e.g., 1, 2, 3, 4,5, 6, 7, 8, 9, 10, 11, 12, 13, 14, 15, 16, 17, 18, 19, or 20 carbonatoms. The alkylene group can be straight, branched or cyclic. Thealkylene group also can be optionally unsaturated and/or substitutedwith one or more “alkyl group substituents.” There can be optionallyinserted along the alkylene group one or more oxygen, sulfur orsubstituted or unsubstituted nitrogen atoms (also referred to herein as“alkylaminoalkyl”), wherein the nitrogen substituent is alkyl aspreviously described. Exemplary alkylene groups include methylene(—CH₂—); ethylene (—CH₂—CH₂—); propylene (—(CH₂)₃—); cyclohexylene(—C₆H₁₀); —CH═CH—CH═CH—; —CH═CH—CH₂—; —CH₂CH₂CH₂CH₂—, —CH₂CH═CHCH₂—,—CH₂CsCCH₂—, —CH₂CH₂CH(CH₂CH₂CH₃)CH₂—, —(CH₂)_(q)—N(R)—(CH₂)_(r)—,wherein each of q and r is independently an integer from 0 to about 20,e.g., 0, 1, 2, 3, 4, 5, 6, 7, 8, 9, 10, 11, 12, 13, 14, 15, 16, 17, 18,19, or 20, and R is hydrogen or lower alkyl; methylenedioxyl(—O—CH₂—O—); and ethylenedioxyl (—O—(CH₂)₂—O—). An alkylene group canhave about 2 to about 3 carbon atoms and can further have 6-20 carbons.Typically, an alkyl (or alkylene) group will have from 1 to 24 carbonatoms, with those groups having 10 or fewer carbon atoms being someembodiments of the present disclosure. A “lower alkyl” or “loweralkylene” is a shorter chain alkyl or alkylene group, generally havingeight or fewer carbon atoms.

The term “heteroalkylene” by itself or as part of another substituentmeans a divalent group derived from heteroalkyl, as exemplified, but notlimited by, —CH₂—CH₂—S—CH₂—CH₂— and —CH₂—S—CH₂—CH₂—NH—CH₂—. Forheteroalkylene groups, heteroatoms also can occupy either or both of thechain termini (e.g., alkyleneoxo, alkylenedioxo, alkyleneamino,alkylenediamino, and the like). Still further, for alkylene andheteroalkylene linking groups, no orientation of the linking group isimplied by the direction in which the formula of the linking group iswritten. For example, the formula —C(O)OR′— represents both —C(O)OR′—and —R′OC(O)—.

The term “aryl” means, unless otherwise stated, an aromatic hydrocarbonsubstituent that can be a single ring or multiple rings (such as from 1to 3 rings), which are fused together or linked covalently. The term“heteroaryl” refers to aryl groups (or rings) that contain from one tofour heteroatoms (in each separate ring in the case of multiple rings)selected from N, O, and S, wherein the nitrogen and sulfur atoms areoptionally oxidized, and the nitrogen atom(s) are optionallyquaternized. A heteroaryl group can be attached to the remainder of themolecule through a carbon or heteroatom. Non-limiting examples of aryland heteroaryl groups include phenyl, 1-naphthyl, 2-naphthyl,4-biphenyl, 1-pyrrolyl, 2-pyrrolyl, 3-pyrrolyl, 3-pyrazolyl,2-imidazolyl, 4-imidazolyl, pyrazinyl, 2-oxazolyl, 4-oxazolyl,2-phenyl-4-oxazolyl, 5-oxazolyl, 3-isoxazolyl, 4-isoxazolyl,5-isoxazolyl, 2-thiazolyl, 4-thiazolyl, 5-thiazolyl, 2-furyl, 3-furyl,2-thienyl, 3-thienyl, 2-pyridyl, 3-pyridyl, 4-pyridyl, 2-pyrimidyl,4-pyrimidyl, 5-benzothiazolyl, purinyl, 2-benzimidazolyl, 5-indolyl,1-isoquinolyl, 5-isoquinolyl, 2-quinoxalinyl, 5-quinoxalinyl,3-quinolyl, and 6-quinolyl. Substituents for each of above noted aryland heteroaryl ring systems are selected from the group of acceptablesubstituents described below. The terms “arylene” and “heteroarylene”refer to the divalent forms of aryl and heteroaryl, respectively.

For brevity, the term “aryl” when used in combination with other terms(e.g., aryloxy, arylthioxy, arylalkyl) includes both aryl and heteroarylrings as defined above. Thus, the terms “arylalkyl” and“heteroarylalkyl” are meant to include those groups in which an aryl orheteroaryl group is attached to an alkyl group (e.g., benzyl, phenethyl,pyridylmethyl, furylmethyl, and the like) including those alkyl groupsin which a carbon atom (e.g., a methylene group) has been replaced by,for example, an oxygen atom (e.g., phenoxymethyl, 2-pyridyloxymethyl,3-(1-naphthyloxy)propyl, and the like). However, the term “haloaryl,” asused herein is meant to cover only aryls substituted with one or morehalogens.

Where a heteroalkyl, heterocycloalkyl, or heteroaryl includes a specificnumber of members (e.g. “3 to 7 membered”), the term “member” refers toa carbon or heteroatom.

Further, a structure represented generally by the formula:

as used herein refers to a ring structure, for example, but not limitedto a 3-carbon, a 4-carbon, a 5-carbon, a 6-carbon, a 7-carbon, and thelike, aliphatic and/or aromatic cyclic compound, including a saturatedring structure, a partially saturated ring structure, and an unsaturatedring structure, comprising a substituent R group, wherein the R groupcan be present or absent, and when present, one or more R groups caneach be substituted on one or more available carbon atoms of the ringstructure. The presence or absence of the R group and number of R groupsis determined by the value of the variable “n,” which is an integergenerally having a value ranging from 0 to the number of carbon atoms onthe ring available for substitution. Each R group, if more than one, issubstituted on an available carbon of the ring structure rather than onanother R group. For example, the structure above where n is 0 to 2would comprise compound groups including, but not limited to:

and the like.

A dashed line representing a bond in a cyclic ring structure indicatesthat the bond can be either present or absent in the ring. That is, adashed line representing a bond in a cyclic ring structure indicatesthat the ring structure is selected from the group consisting of asaturated ring structure, a partially saturated ring structure, and anunsaturated ring structure.

The symbol (

) denotes the point of attachment of a moiety to the remainder of themolecule.

When a named atom of an aromatic ring or a heterocyclic aromatic ring isdefined as being “absent,” the named atom is replaced by a direct bond.

Each of above terms (e.g., “alkyl,” “heteroalkyl,” “cycloalkyl, and“heterocycloalkyl”, “aryl,” “heteroaryl,” “phosphonate,” and “sulfonate”as well as their divalent derivatives) are meant to include bothsubstituted and unsubstituted forms of the indicated group. Optionalsubstituents for each type of group are provided below.

Substituents for alkyl, heteroalkyl, cycloalkyl, heterocycloalkylmonovalent and divalent derivative groups (including those groups oftenreferred to as alkylene, alkenyl, heteroalkylene, heteroalkenyl,alkynyl, cycloalkyl, heterocycloalkyl, cycloalkenyl, andheterocycloalkenyl) can be one or more of a variety of groups selectedfrom, but not limited to: —OR′, ═O, ═NR′, ═NR—OR′, —NR′R″, —SR′,-halogen, —SiR′R″R′″, —OC(O)R′, —C(O)R′, —CO₂R′, —C(O)NR′R″,—OC(O)NR′R″, —NR″C(O)R′, —NR′—C(O)NR″R′″, —NR″C(O)OR′,—NR—C(NR′R″)═NR′″, —S(O)R′, —S(O)₂R′, —S(O)₂NR′R″, —NRSO₂R′, —CN and—NO₂ in a number ranging from zero to (2m′+1), where m′ is the totalnumber of carbon atoms in such groups. R′, R″, R′″ and R″″ each mayindependently refer to hydrogen, substituted or unsubstitutedheteroalkyl, substituted or unsubstituted cycloalkyl, substituted orunsubstituted heterocycloalkyl, substituted or unsubstituted aryl (e.g.,aryl substituted with 1-3 halogens), substituted or unsubstituted alkyl,alkoxy or thioalkoxy groups, or arylalkyl groups. As used herein, an“alkoxy” group is an alkyl attached to the remainder of the moleculethrough a divalent oxygen. When a compound of the disclosure includesmore than one R group, for example, each of the R groups isindependently selected as are each R′, R″, R′″ and R″″ groups when morethan one of these groups is present. When R′ and R″ are attached to thesame nitrogen atom, they can be combined with the nitrogen atom to forma 4-, 5-, 6-, or 7-membered ring. For example, —NR′R″ is meant toinclude, but not be limited to, 1-pyrrolidinyl and 4-morpholinyl. Fromthe above discussion of substituents, one of skill in the art willunderstand that the term “alkyl” is meant to include groups includingcarbon atoms bound to groups other than hydrogen groups, such ashaloalkyl (e.g., —CF₃ and —CH₂CF₃) and acyl (e.g., —C(O)CH₃, —C(O)CF₃,—C(O)CH₂OCH₃, and the like).

Similar to the substituents described for alkyl groups above, exemplarysubstituents for aryl and heteroaryl groups (as well as their divalentderivatives) are varied and are selected from, for example: halogen,—OR′, —NR′R″, —SR′, —SiR′R″R′″, —OC(O)R′, —C(O)R′, —CO₂R′, —C(O)NR′R″,—OC(O)NR′R″, —NR″C(O)R′, —NR′—C(O)NR″R′″, —NR″C(O)OR′,—NR—C(NR′R″R′″)═NR″″, —NR—C(NR′R″)═NR′″—S(O)R′, —S(O)₂R′, —S(O)₂NR′R″,—NRSO₂R′, —CN and —NO₂, —R′, —N₃, —CH(Ph)₂, fluoro(C₁-C₄)alkoxo, andfluoro(C₁-C₄)alkyl, in a number ranging from zero to the total number ofopen valences on aromatic ring system; and where R′, R″, R′″ and R″″ maybe independently selected from hydrogen, substituted or unsubstitutedalkyl, substituted or unsubstituted heteroalkyl, substituted orunsubstituted cycloalkyl, substituted or unsubstituted heterocycloalkyl,substituted or unsubstituted aryl and substituted or unsubstitutedheteroaryl. When a compound of the disclosure includes more than one Rgroup, for example, each of the R groups is independently selected asare each R′, R″, R′″ and R″″ groups when more than one of these groupsis present.

Two of the substituents on adjacent atoms of aryl or heteroaryl ring mayoptionally form a ring of the formula -T-C(O)—(CRR′)_(q)—U—, wherein Tand U are independently —NR—, —O—, —CRR′— or a single bond, and q is aninteger of from 0 to 3. Alternatively, two of the substituents onadjacent atoms of aryl or heteroaryl ring may optionally be replacedwith a substituent of the formula -A-(CH₂)_(r)—B—, wherein A and B areindependently —CRR′—, —O—, —NR—, —S—, —S(O)—, —S(O)₂—, —S(O)₂NR′— or asingle bond, and r is an integer of from 1 to 4.

One of the single bonds of the new ring so formed may optionally bereplaced with a double bond. Alternatively, two of the substituents onadjacent atoms of aryl or heteroaryl ring may optionally be replacedwith a substituent of the formula —(CRR′)_(s)—X′—(C″R′″)_(d)—, where sand d are independently integers of from 0 to 3, and X′ is —O—, —NR′—,—S—, —S(O)—, —S(O)₂—, or —S(O)₂NR′—. The substituents R, R′, R″ and R′″may be independently selected from hydrogen, substituted orunsubstituted alkyl, substituted or unsubstituted cycloalkyl,substituted or unsubstituted heterocycloalkyl, substituted orunsubstituted aryl, and substituted or unsubstituted heteroaryl.

As used herein, the term “acyl” refers to an organic acid group whereinthe —OH of the carboxyl group has been replaced with another substituentand has the general formula RC(═O)—, wherein R is an alkyl, alkenyl,alkynyl, aryl, carbocylic, heterocyclic, or aromatic heterocyclic groupas defined herein). As such, the term “acyl” specifically includesarylacyl groups, such as a 2-(furan-2-yl)acetyl)- and a 2-phenylacetylgroup. Specific examples of acyl groups include acetyl and benzoyl. Acylgroups also are intended to include amides, —RC(═O)NR′, esters,—RC(═O)OR′, ketones, —RC(═O)R′, and aldehydes, —RC(═O)H.

The terms “alkoxyl” or “alkoxy” are used interchangeably herein andrefer to a saturated (i.e., alkyl-O—) or unsaturated (i.e., alkenyl-O—and alkynyl-O—) group attached to the parent molecular moiety through anoxygen atom, wherein the terms “alkyl,” “alkenyl,” and “alkynyl” are aspreviously described and can include C₁₋₂₀ inclusive, linear, branched,or cyclic, saturated or unsaturated oxo-hydrocarbon chains, including,for example, methoxyl, ethoxyl, propoxyl, isopropoxyl, n-butoxyl,sec-butoxyl, tert-butoxyl, and n-pentoxyl, neopentoxyl, n-hexoxyl, andthe like.

The term “alkoxyalkyl” as used herein refers to an alkyl-O-alkyl ether,for example, a methoxyethyl or an ethoxymethyl group.

“Aryloxyl” refers to an aryl-O— group wherein the aryl group is aspreviously described, including a substituted aryl. The term “aryloxyl”as used herein can refer to phenyloxyl or hexyloxyl, and alkyl,substituted alkyl, halo, or alkoxyl substituted phenyloxyl or hexyloxyl.

“Aralkyl” refers to an aryl-alkyl-group wherein aryl and alkyl are aspreviously described, and included substituted aryl and substitutedalkyl. Exemplary aralkyl groups include benzyl, phenylethyl, andnaphthylmethyl.

“Aralkyloxyl” refers to an aralkyl-O— group wherein the aralkyl group isas previously described. An exemplary aralkyloxyl group is benzyloxyl,i.e., C₆H₅—CH₂—O—. An aralkyloxyl group can optionally be substituted.

“Alkoxycarbonyl” refers to an alkyl-O—C(═O)— group. Exemplaryalkoxycarbonyl groups include methoxycarbonyl, ethoxycarbonyl,butyloxycarbonyl, and tert-butyloxycarbonyl.

“Aryloxycarbonyl” refers to an aryl-O—C(═O)— group. Exemplaryaryloxycarbonyl groups include phenoxy- and naphthoxy-carbonyl.

“Aralkoxycarbonyl” refers to an aralkyl-O—C(═O)— group. An exemplaryaralkoxycarbonyl group is benzyloxycarbonyl.

“Carbamoyl” refers to an amide group of the formula —C(═O)NH₂.“Alkylcarbamoyl” refers to a R′RN—C(═O)— group wherein one of R and R′is hydrogen and the other of R and R′ is alkyl and/or substituted alkylas previously described. “Dialkylcarbamoyl” refers to a R′RN—C(═O)—group wherein each of R and R′ is independently alkyl and/or substitutedalkyl as previously described.

The term carbonyldioxyl, as used herein, refers to a carbonate group ofthe formula —O—C(═O)—OR.

“Acyloxyl” refers to an acyl-O— group wherein acyl is as previouslydescribed.

The term “amino” refers to the —NH₂ group and also refers to a nitrogencontaining group as is known in the art derived from ammonia by thereplacement of one or more hydrogen radicals by organic radicals. Forexample, the terms “acylamino” and “alkylamino” refer to specificN-substituted organic radicals with acyl and alkyl substituent groupsrespectively.

An “aminoalkyl” as used herein refers to an amino group covalently boundto an alkylene linker. More particularly, the terms alkylamino,dialkylamino, and trialkylamino as used herein refer to one, two, orthree, respectively, alkyl groups, as previously defined, attached tothe parent molecular moiety through a nitrogen atom. The term alkylaminorefers to a group having the structure —NHR′ wherein R′ is an alkylgroup, as previously defined; whereas the term dialkylamino refers to agroup having the structure —NR′R wherein R′ and R″ are eachindependently selected from the group consisting of alkyl groups. Theterm trialkylamino refers to a group having the structure —NR′R″R′″,wherein R′, R″, and R′″ are each independently selected from the groupconsisting of alkyl groups. Additionally, R′, R″, and/or R′″ takentogether may optionally be —(CH₂)_(k)— where k is an integer from 2 to6. Examples include, but are not limited to, methylamino, dimethylamino,ethylamino, diethylamino, diethylaminocarbonyl, methylethylamino,isopropylamino, piperidino, trimethylamino, and propylamino.

The amino group is —NR′R″, wherein R′ and R″ are typically selected fromhydrogen, substituted or unsubstituted alkyl, substituted orunsubstituted heteroalkyl, substituted or unsubstituted cycloalkyl,substituted or unsubstituted heterocycloalkyl, substituted orunsubstituted aryl, or substituted or unsubstituted heteroaryl.

The terms alkylthioether and thioalkoxyl refer to a saturated (i.e.,alkyl-S—) or unsaturated (i.e., alkenyl-S— and alkynyl-S—) groupattached to the parent molecular moiety through a sulfur atom. Examplesof thioalkoxyl moieties include, but are not limited to, methylthio,ethylthio, propylthio, isopropylthio, n-butylthio, and the like.

“Acylamino” refers to an acyl-NH— group wherein acyl is as previouslydescribed. “Aroylamino” refers to an aroyl-NH— group wherein aroyl is aspreviously described.

The term “carbonyl” refers to the —C(═O)— group, and can include analdehyde group represented by the general formula R—C(═O)H.

The term “carboxyl” refers to the —COOH group. Such groups also arereferred to herein as a “carboxylic acid” moiety.

The terms “halo,” “halide,” or “halogen” as used herein refer to fluoro,chloro, bromo, and iodo groups. Additionally, terms such as “haloalkyl,”are meant to include monohaloalkyl and polyhaloalkyl. For example, theterm “halo(C₁-C₄)alkyl” is mean to include, but not be limited to,trifluoromethyl, 2,2,2-trifluoroethyl, 4-chlorobutyl, 3-bromopropyl, andthe like.

The term “hydroxyl” refers to the —OH group.

The term “hydroxyalkyl” refers to an alkyl group substituted with an —OHgroup.

The term “mercapto” refers to the —SH group.

The term “oxo” as used herein means an oxygen atom that is double bondedto a carbon atom or to another element.

The term “nitro” refers to the —NO₂ group.

The term “thio” refers to a compound described previously herein whereina carbon or oxygen atom is replaced by a sulfur atom.

The term “sulfate” refers to the —SO₄ group.

The term thiohydroxyl or thiol, as used herein, refers to a group of theformula —SH.

More particularly, the term “sulfide” refers to compound having a groupof the formula —SR.

The term “sulfone” refers to compound having a sulfonyl group —S(O₂)R.

The term “sulfoxide” refers to a compound having a sulfinyl group —S(O)R

The term ureido refers to a urea group of the formula —NH—CO—NH₂.

Throughout the specification and claims, a given chemical formula orname shall encompass all tautomers, congeners, and optical- andstereoisomers, as well as racemic mixtures where such isomers andmixtures exist.

Certain compounds of the present disclosure may possess asymmetriccarbon atoms (optical or chiral centers) or double bonds; theenantiomers, racemates, diastereomers, tautomers, geometric isomers,stereoisometric forms that may be defined, in terms of absolutestereochemistry, as (R)- or (S)- or, as D- or L- for amino acids, andindividual isomers are encompassed within the scope of the presentdisclosure. The compounds of the present disclosure do not include thosewhich are known in art to be too unstable to synthesize and/or isolate.The present disclosure is meant to include compounds in racemic,scalemic, and optically pure forms. Optically active (R)- and (S)-, orD- and L-isomers may be prepared using chiral synthons or chiralreagents, or resolved using conventional techniques. When the compoundsdescribed herein contain olefenic bonds or other centers of geometricasymmetry, and unless specified otherwise, it is intended that thecompounds include both E and Z geometric isomers. Unless otherwisestated, structures depicted herein are also meant to include allstereochemical forms of the structure; i.e., the R and S configurationsfor each asymmetric center. Therefore, single stereochemical isomers aswell as enantiomeric and diastereomeric mixtures of the presentcompounds are within the scope of the disclosure.

It will be apparent to one skilled in the art that certain compounds ofthis disclosure may exist in tautomeric forms, all such tautomeric formsof the compounds being within the scope of the disclosure. The term“tautomer,” as used herein, refers to one of two or more structuralisomers which exist in equilibrium and which are readily converted fromone isomeric form to another.

Unless otherwise stated, structures depicted herein are also meant toinclude compounds which differ only in the presence of one or moreisotopically enriched atoms. For example, compounds having the presentstructures with the replacement of a hydrogen by a deuterium or tritium,or the replacement of a carbon by ¹³C- or ¹⁴C-enriched carbon are withinthe scope of this disclosure.

The compounds of the present disclosure may also contain unnaturalproportions of atomic isotopes at one or more of atoms that constitutesuch compounds. For example, the compounds may be radiolabeled withradioactive isotopes, such as for example iodine-125 (¹²⁵I) orastatine-211 (²¹¹At). All isotopic variations of the compounds of thepresent disclosure, whether radioactive or not, are encompassed withinthe scope of the present disclosure.

The compounds of the present disclosure may exist as salts. The presentdisclosure includes such salts. Examples of applicable salt formsinclude hydrochlorides, hydrobromides, sulfates, methanesulfonates,nitrates, maleates, acetates, citrates, fumarates, tartrates (e.g.(+)-tartrates, (−)-tartrates or mixtures thereof including racemicmixtures, succinates, benzoates and salts with amino acids such asglutamic acid. These salts may be prepared by methods known to thoseskilled in art. Also included are base addition salts such as sodium,potassium, calcium, ammonium, organic amino, or magnesium salt, or asimilar salt. When compounds of the present disclosure containrelatively basic functionalities, acid addition salts can be obtained bycontacting the neutral form of such compounds with a sufficient amountof the desired acid, either neat or in a suitable inert solvent or byion exchange. Examples of acceptable acid addition salts include thosederived from inorganic acids like hydrochloric, hydrobromic, nitric,carbonic, monohydrogencarbonic, phosphoric, monohydrogenphosphoric,dihydrogenphosphoric, sulfuric, monohydrogensulfuric, hydriodic, orphosphorous acids and the like, as well as the salts derived organicacids like acetic, propionic, isobutyric, maleic, malonic, benzoic,succinic, suberic, fumaric, lactic, mandelic, phthalic, benzenesulfonic,p-tolylsulfonic, citric, tartaric, methanesulfonic, and the like. Alsoincluded are salts of amino acids such as arginate and the like, andsalts of organic acids like glucuronic or galactunoric acids and thelike. Certain specific compounds of the present disclosure contain bothbasic and acidic functionalities that allow the compounds to beconverted into either base or acid addition salts.

The neutral forms of the compounds may be regenerated by contacting thesalt with a base or acid and isolating the parent compound in theconventional manner. The parent form of the compound differs from thevarious salt forms in certain physical properties, such as solubility inpolar solvents.

Certain compounds of the present disclosure can exist in unsolvatedforms as well as solvated forms, including hydrated forms. In general,the solvated forms are equivalent to unsolvated forms and areencompassed within the scope of the present disclosure. Certaincompounds of the present disclosure may exist in multiple crystalline oramorphous forms. In general, all physical forms are equivalent for theuses contemplated by the present disclosure and are intended to bewithin the scope of the present disclosure.

In addition to salt forms, the present disclosure provides compounds,which are in a prodrug form. Prodrugs of the compounds described hereinare those compounds that readily undergo chemical changes underphysiological conditions to provide the compounds of the presentdisclosure. Additionally, prodrugs can be converted to the compounds ofthe present disclosure by chemical or biochemical methods in an ex vivoenvironment. For example, prodrugs can be slowly converted to thecompounds of the present disclosure when placed in a transdermal patchreservoir with a suitable enzyme or chemical reagent.

The term “protecting group” refers to chemical moieties that block someor all reactive moieties of a compound and prevent such moieties fromparticipating in chemical reactions until the protective group isremoved, for example, those moieties listed and described in T. W.Greene, P.G.M. Wuts, Protective Groups in Organic Synthesis, 3rd ed.John Wiley & Sons (1999). It may be advantageous, where differentprotecting groups are employed, that each (different) protective groupbe removable by a different means. Protective groups that are cleavedunder totally disparate reaction conditions allow differential removalof such protecting groups. For example, protective groups can be removedby acid, base, and hydrogenolysis. Groups such as trityl,dimethoxytrityl, acetal and tert-butyldimethylsilyl are acid labile andmay be used to protect carboxy and hydroxy reactive moieties in thepresence of amino groups protected with Cbz groups, which are removableby hydrogenolysis, and Fmoc groups, which are base labile. Carboxylicacid and hydroxy reactive moieties may be blocked with base labilegroups such as, without limitation, methyl, ethyl, and acetyl in thepresence of amines blocked with acid labile groups such as tert-butylcarbamate or with carbamates that are both acid and base stable buthydrolytically removable.

Carboxylic acid and hydroxy reactive moieties may also be blocked withhydrolytically removable protective groups such as the benzyl group,while amine groups capable of hydrogen bonding with acids may be blockedwith base labile groups such as Fmoc. Carboxylic acid reactive moietiesmay be blocked with oxidatively-removable protective groups such as2,4-dimethoxybenzyl, while co-existing amino groups may be blocked withfluoride labile silyl carbamates.

Allyl blocking groups are useful in the presence of acid- andbase-protecting groups since the former are stable and can besubsequently removed by metal or pi-acid catalysts. For example, anallyl-blocked carboxylic acid can be deprotected with a palladium(O)—catalyzed reaction in the presence of acid labile t-butyl carbamate orbase-labile acetate amine protecting groups. Yet another form ofprotecting group is a resin to which a compound or intermediate may beattached. As long as the residue is attached to the resin, thatfunctional group is blocked and cannot react. Once released from theresin, the functional group is available to react.

Typical blocking/protecting groups include, but are not limited to thefollowing moieties:

Further, as used herein, a “protecting group” is a chemical substituentwhich can be selectively removed by readily available reagents which donot attack the regenerated functional group or other functional groupsin the molecule. Suitable protecting groups are known in the art andcontinue to be developed. Suitable protecting groups may be found, forexample in Wutz et al. (“Greene's Protective Groups in OrganicSynthesis, Fourth Edition,” Wiley-Interscience, 2007). Protecting groupsfor protection of the carboxyl group, as described by Wutz et al. (pages533-643), are used in certain embodiments. In some embodiments, theprotecting group is removable by treatment with acid. Specific examplesof protecting groups include, but are not limited to, benzyl,p-methoxybenzyl (PMB), tertiary butyl (t-Bu), methoxymethyl (MOM),methoxyethoxymethyl (MEM), methylthiomethyl (MTM), tetrahydropyranyl(THP), tetrahydrofuranyl (THF), benzyloxymethyl (BOM), trimethylsilyl(TMS), triethylsilyl (TES), t-butyldimethylsilyl (TBDMS), andtriphenylmethyl (trityl, Tr). Persons skilled in the art will recognizeappropriate situations in which protecting groups are required and willbe able to select an appropriate protecting group for use in aparticular circumstance.

The term “metal ion” as used herein refers to elements of the periodictable that are metallic and that are positively charged as a result ofhaving fewer electrons in the valence shell than is present for theneutral metallic element. Metals that are useful in the presentlydisclosed subject matter include metals capable of formingpharmaceutically acceptable compositions. Useful metals include, but arenot limited to, Li, Na, K, Rb, Cs, Be, Mg, Ca, Sr, and Ba. One of skillin the art will appreciate that the metals described above can eachadopt several different oxidation states. In some instances, the moststable oxidation state is formed, but other oxidation states are usefulin the presently disclosed subject matter.

Following long-standing patent law convention, the terms “a,” “an,” and“the” refer to “one or more” when used in this application, includingthe claims. Thus, for example, reference to “a subject” includes aplurality of subjects, unless the context clearly is to the contrary(e.g., a plurality of subjects), and so forth.

Throughout this specification and the claims, the terms “comprise,”“comprises,” and “comprising” are used in a non-exclusive sense, exceptwhere the context requires otherwise. Likewise, the term “include” andits grammatical variants are intended to be non-limiting, such thatrecitation of items in a list is not to the exclusion of other likeitems that can be substituted or added to the listed items.

For the purposes of this specification and appended claims, unlessotherwise indicated, all numbers expressing amounts, sizes, dimensions,proportions, shapes, formulations, parameters, percentages, quantities,characteristics, and other numerical values used in the specificationand claims, are to be understood as being modified in all instances bythe term “about” even though the term “about” may not expressly appearwith the value, amount or range. Accordingly, unless indicated to thecontrary, the numerical parameters set forth in the followingspecification and attached claims are not and need not be exact, but maybe approximate and/or larger or smaller as desired, reflectingtolerances, conversion factors, rounding off, measurement error and thelike, and other factors known to those of skill in the art depending onthe desired properties sought to be obtained by the presently disclosedsubject matter. For example, the term “about,” when referring to a valuecan be meant to encompass variations of, in some embodiments, ±100% insome embodiments ±50%, in some embodiments ±20%, in some embodiments±10%, in some embodiments ±5%, in some embodiments ±1%, in someembodiments ±0.5%, and in some embodiments ±0.1% from the specifiedamount, as such variations are appropriate to perform the disclosedmethods or employ the disclosed compositions.

Further, the term “about” when used in connection with one or morenumbers or numerical ranges, should be understood to refer to all suchnumbers, including all numbers in a range and modifies that range byextending the boundaries above and below the numerical values set forth.The recitation of numerical ranges by endpoints includes all numbers,e.g., whole integers, including fractions thereof, subsumed within thatrange (for example, the recitation of 1 to 5 includes 1, 2, 3, 4, and 5,as well as fractions thereof, e.g., 1.5, 2.25, 3.75, 4.1, and the like)and any range within that range.

In the examples below the following terms are intended to have thefollowing meaning: ACN: acetonitrile, DCM: Dichloromethane, DIPEA:N,N-Diisopropylethylamine, DMF: Dimethylformamide, HPLC: HighPerformance Liquid Chromatography, HRMS: High Resolution MassSpectrometry, LRMS: Low Resolution Mass Spectrometry, NCS:N-Chlorosuccinimide, NHS: N-Hydroxysuccinimide, NMR: nuclear magneticresonance, PMB: p-methoxybenzyl, RT: room temperature, TEA:Triethylamine, TFA: Trifluoroacetic acid, and TSTU:O—(N-Succinimidyl)-N,N,N′,N′-tetramethyluronium tetrafluoroborate.

EXAMPLES

The following Examples have been included to provide guidance to one ofordinary skill in the art for practicing representative embodiments ofthe presently disclosed subject matter. In light of the presentdisclosure and the general level of skill in the art, those of skill canappreciate that the following Examples are intended to be exemplary onlyand that numerous changes, modifications, and alterations can beemployed without departing from the scope of the presently disclosedsubject matter. The synthetic descriptions and specific examples thatfollow are only intended for the purposes of illustration, and are notto be construed as limiting in any manner to make compounds of thedisclosure by other methods.

Example 1 General Methods

Chemistry.

All chemicals and solvents were purchased from either Sigma-Aldrich(Milwaukee, Wis.) or Fisher Scientific (Pittsburgh, Pa.). TheN-hydroxysuccinimide (NHS) esters of DyLight 800 was purchased fromThermo Fisher Scientific (Rockford, Ill.). ESI mass spectra wereobtained on a Bruker Esquire 3000 plus system (Billerica, Mass.).High-performance liquid chromatography (HPLC) purifications wereperformed on a Varian Prostar System (Varian Medical Systems, Palo Alto,Calif.).

Cell Lines and Tumor Models.

PSMA⁺ PC3 PIP and PSMA⁻ PC3 flu cell lines were obtained from Dr. WarrenHeston (Cleveland Clinic). Cells were grown to 80-90% confluence in asingle passage before trypsinization and formulation in Hank's balancedsalt solution (HBSS, Sigma, St. Louis, Mo.) for implantation into mice.Animal studies were carried out in compliance with guidelines related tothe conduct of animal experiments of the Johns Hopkins Animal Care andUse Committee. For optical imaging studies and ex-vivo biodistribution,male NOD-SCID mice (JHU, in house colony) were implanted subcutaneouslywith 1×10⁶ PSMA⁺ PC3 PIP and PSMA⁻ PC3 flu cells in opposite flanks.Mice were imaged when the tumor xenografts reached 3-5 mm in diameter.

In Vivo Imaging and Ex Vivo Biodistribution.

After image acquisition at baseline (pre-injection), mouse was injectedintravenously with 1 nmol of DyLight800-3 and images were acquired at 1h, 2 h, 4 h and 24 h time points using a Pearl Impulse Imager (LI-CORBiosciences). Following the 24 h image the mouse was sacrificed bycervical dislocation and tumor, muscle, liver, spleen, kidneys andintestine were collected and assembled on a petri dish for imageacquisition. All images were scaled to the same intensity for directcomparison. FIG. 1 shows the images at 24 hours postinjection of 1 nmolof DyLight800-3 in mouse with PSMA+ PC3 PIP and PSMA− PC3 flu tumors.Both whole body and ex vivo organ imaging clearly demonstrated PSMA+ PC3PIP tumor uptake and little uptake in PSMA− PC3 flu tumor, indicatingtarget selectivity in vivo.

Example 2 Synthesis Methods

Synthesis of DyLight800-3:

To a solution of compound 3, Chen et al., 2009, (0.5 mg, 0.7 μmol) inDMSO (0.1 mL) was added N,N-diisopropylethylamine (0.010 mL, 57.4 μmol),followed by NHS ester of DyLight800 (0.3 mg, 0.29 μmol). After 1 h atroom temperature, the reaction mixture was purified by HPLC (column,Phenomenex Luna C18 10μ, 250×4.6 mm; mobile phase, A=0.1% TFA in H₂O,B=0.1% TFA in CH₃CN; gradient, 0 min=5% B, 5 min=5% B, 45 min=100% B;flow rate, 1 mL/min) to afford 0.3 mg (70%) of DyLight800-3: ESI-Masscalcd for C₇₁H₉₄N₇O₂₂S₃ ⁻ [M-H]⁻ 1492.6, found 1492.4 [M-H]⁻.

REFERENCES

All publications, patent applications, patents, and other referencesmentioned in the specification are indicative of the level of thoseskilled in the art to which the presently disclosed subject matterpertains. All publications, patent applications, patents, and otherreferences are herein incorporated by reference to the same extent as ifeach individual publication, patent application, patent, and otherreference was specifically and individually indicated to be incorporatedby reference. It will be understood that, although a number of patentapplications, patents, and other references are referred to herein, suchreference does not constitute an admission that any of these documentsforms part of the common general knowledge in the art. In case of aconflict between the specification and any of the incorporatedreferences, the specification (including any amendments thereof, whichmay be based on an incorporated reference), shall control. Standardart-accepted meanings of terms are used herein unless indicatedotherwise. Standard abbreviations for various terms are used herein.

-   Chen Y, Pullambhatla M, Banerjee S, Byun Y, Stathis M, Rojas C,    Slusher B S, Mease R C, Pomper M G. Synthesis and biological    evaluation of low molecular weight fluorescent imaging agents for    the prostate-specific membrane antigen. Bioconjug Chem. 23: 2377-85    (2012);-   Maresca K P, Hillier S M, Femia F J, Keith D, Barone C, Joyal J L,    Zimmerman C N, Kozikowski A P, Barrett J A, Eckelman W C, Babic J W.    A Series of Halogenated Heterodimeric Inhibitors of Prostate    Specific Membrane Antigen (PSMA) as Radiolabeled Probes for    Targeting Prostate Cancer J. Med. Chem. 52: 347-357 (2009);-   Chen Y, Dhara S, Banerjee S, Byun Y, Pullambhatla M, Mease R C,    Pomper M G. A low molecular weight PSMA-based fluorescent imaging    agent for cancer. Biochem. Biophys Res. Commun. 390: 624-629 (2009);-   Pomper, Martin G.; Mease, Ronnie C.; Ray, Sangeeta; Chen, Ying    Psma-targeting compounds and uses thereof;-   International PCT patent application publication no.    WO2010/108125A2, for PSMA-TARGETING COMPOUNDS AND USES THEREOF, to    Pomper et al., published Sep. 23, 2010.-   Rowe, S P, Gorin M S, Hammers H J, Javadi M S, Hawasli H, Szabo Z,    Cho S Y, Pomper M G, Allaf M E. Imaging of metastatic clear cell    renal cell carcinoma with PSMA-targeted ¹⁸F-DCFPyL PET/CT. Ann.    Nucl. Med. 29(10) 877-882 2015.

Although the foregoing subject matter has been described in some detailby way of illustration and example for purposes of clarity ofunderstanding, it will be understood by those skilled in the art thatcertain changes and modifications can be practiced within the scope ofthe appended claims.

That which is claimed:
 1. A compound of the following formula:

or a pharmaceutically acceptable salt thereof.
 2. A compositioncomprising compound (3):

or a pharmaceutically acceptable salt thereof; wherein the compositionis adapted for visualization of tissue under illumination withnear-infrared radiation.
 3. The composition of claim 2, wherein thecomposition is adapted for administration to a subject.
 4. Thecomposition of claim 3, wherein the composition comprises a unit dosageform of compound (3).
 5. The composition of claim 4, wherein the unitdosage form delivers to the subject an amount of compound (3) betweenabout 0.01 and about 8 mg/kg.
 6. The composition of claim 5, wherein theunit dosage form delivers to the subject an amount of compound (3) ofabout 0.01 mg/kg, about 0.05 mg/kg, about 0.10 mg/kg, about 0.20 mg/kg,about 0.3 mg/kg, about 0.35 mg/kg, about 0.40 mg/kg, about 0.45 mg/kg,about 0.50 mg/kg, about 0.55 mg/kg, about 0.60 mg/kg, about 0.65 mg/kg,about 0.70 mg/kg, about 0.75 mg/kg, about 0.80 mg/kg, about 0.90 mg/kg,about 1 mg/kg, about 2, mg/kg, about 4 mg/kg, about 6 mg/kg, or about 8mg/kg.
 7. The composition of claim 2, wherein the composition is in asingle dose form.
 8. The composition of claim 2, wherein the compositionis in dry form.
 9. The composition of claim 2, wherein the compositionis lyophilized in a sterile container.
 10. The composition of claim 2,wherein the composition is contained within a sterile container.
 11. Thecomposition of claim 10, wherein the sterile container comprises amachine detectable identifier.
 12. The composition of claim 2, furthercomprising one or more pharmaceutically acceptable excipients in an oraldosage form.
 13. The composition of claim 2, further comprising one ormore pharmaceutically acceptable carriers in an injectable dosage form.14. The composition of claim 2, further comprising one or morepharmaceutically acceptable excipients in a dosage form for directdelivery to a surgical site.
 15. The use of a composition of claim 2 foradministration to a subject to obtain visualization of tissue expressingPSMA under illumination with near-infrared radiation.
 16. The use ofclaim 15, wherein the subject is a human subject.
 17. A method forvisualization of tissue expressing PSMA, the method comprisingadministering to a subject a composition comprising compound (3):

or a pharmaceutically acceptable salt thereof; wherein compound (3) isadministered in an amount sufficient for imaging tissue underillumination with near-infrared radiation; imaging the tissue underillumination with near-infrared radiation; and obtaining at least oneimage of tissue from the subject.
 18. The method of claim 17, whereinthe composition comprises a unit dosage form of compound (3).
 19. Themethod of claim 18, wherein the unit dosage form delivers to the subjectan amount of compound (3) from about 0.01 mg/kg and about 8 mg/kg. 20.The method of claim 18, wherein the composition is sterile, non-toxic,and adapted for administration to a subject.
 21. The method of claim 17,further comprising obtaining the image during administration, afteradministration, or both during and after administration of thecomposition.
 22. The method of claim 17, further comprisingintravenously injecting a composition comprising compound (3) into thesubject.
 23. The method of claim 22, wherein the composition is injectedinto a circulatory system of the subject.
 24. The method of claim 17,further comprising visualizing a subject area on which surgery is orwill be performed.
 25. The method of claim 24, further comprisingperforming a surgical procedure of the subject area based on thevisualization of the area.
 26. The method of claim 24, furthercomprising viewing a subject area on which an ophthalmic, arthroscopic,laparoscopic, cardiothoracic, muscular, or neurological procedure is orwill be performed.
 27. The method of claim 17, further comprisingdiagnosing the subject with a condition or disease based on thevisualization of the tissue expressing PSMA.
 28. The method of claim 17,further comprising obtaining ex vivo images of at least a portion of thesubject.
 29. The method of claim 17, wherein the tissue being visualizedcomprises tumor tissue.
 30. The method of claim 17, wherein the tissuebeing visualized comprises cancerous tissue.
 31. The method of claim 17,wherein the tissue being visualized comprises prostate tissue.
 32. Themethod of claim 17, wherein the tissue being visualized comprisesprostate tumor tissue.
 33. The method of claim 17, wherein the tissuebeing visualized comprises nerve tissue.
 34. The method of claim 17wherein the tissue being visualized comprises clear cell renal cellcarcinoma.